Chronic Alcohol Reduces Bone Mass Through Inhibiting Proliferation and Promoting Aging of Endothelial Cells in Type-H Vessels

Ao Chen; Xiaoting Li; Jingyu Zhao; Jiawen Zhou; Chunfeng Xie; Haiyun Chen; Qiuyi Wang; Rong Wang; Dengshun Miao; Jie Li; Jianliang Jin

doi:10.1089/scd.2021.0337

Chronic Alcohol Reduces Bone Mass Through Inhibiting Proliferation and Promoting Aging of Endothelial Cells in Type-H Vessels

Stem Cells Dev. 2022 Sep;31(17-18):541-554. doi: 10.1089/scd.2021.0337. Epub 2022 Aug 22.

Authors

Ao Chen¹, Xiaoting Li², Jingyu Zhao¹, Jiawen Zhou¹, Chunfeng Xie², Haiyun Chen³, Qiuyi Wang¹, Rong Wang¹, Dengshun Miao^{1

3}, Jie Li⁴, Jianliang Jin¹

Affiliations

¹ Department of Human Anatomy, Research Center for Bone and Stem Cells; Key Laboratory for Aging & Disease, The State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, Jiangsu, China.
² Department of Nutrition and Food Safety, School of Public Health, Nanjing Medical University, Nanjing, Jiangsu, China.
³ The Research Center for Aging, Affiliated Friendship Plastic Surgery Hospital of Nanjing Medical University, Nanjing, China.
⁴ Department of Orthopedics, Xuzhou Central Hospital, The Xuzhou Clinical School of Xuzhou Medical University, The Xuzhou School of Clinical Medicine of Nanjing Medical University, Xuzhou, China.

PMID: 35491665
DOI: 10.1089/scd.2021.0337

Abstract

Alcohol consumption is regarded as one of the leading risk factors for secondary osteopenia. Angiogenesis and osteogenesis coupled by type-H vessels coordinate the biological process of bone homeostasis to prevent osteopenia. This study aimed to determine whether chronic alcohol inhibits type-H vessel-dependent bone formation. Two-month-old mice were fed with 5% (v/v) alcohol liquid diet (28% of calories) or normal liquid diet every day for 2 months. The tibias were isolated and detected with X-ray and microcomputed tomography. Paraffin-embedded or frozen tibial sections were prepared and used for immunohistochemical or immunofluorescence staining, respectively. Human umbilical vein endothelial cells (HUVECs) were treated with different concentrations of alcohol, including 0 mM (0%), 8.7 mM (0.5%), 52 mM (3%), or 87 mM (5%) alcohol for 12 h. The conditioned medium of the above HUVEC cells was collected to culture human bone marrow-mesenchymal stem cells (BM-MSCs), which were induced to differentiate into osteoblasts in vitro. The alcoholic diet retarded the bone growth and led to osteoporosis, impaired bone formation of osteoblasts, and decreased CD31^hiEMCN^hi type-H vessel formation through inhibiting proliferation and promoting aging of endothelial cells in mice. Alcohol treatment obviously increased the expression of p16, while significantly decreased the expression of Bmi-1, CDK6, Cyclin D, E2F1, and bone morphogenetic protein (BMP)2 compared with vehicle. Alcohol inhibited the differentiation of BM-MSCs into osteoblasts through reducing the BMP2 secretion of endothelial cells in type-H vessels. Alcoholic diet impaired CD31^hiEMCN^hi type-H vessel formation through inhibiting proliferation and promoting aging of endothelial cells through Bmi-1/p16 signaling, and inhibited the differentiation of BM-MSCs into osteoblasts through reducing the BMP2 secretion of endothelial cells in type-H vessels. This study provides a basis for developing a new treatment strategy targeting aging endothelial cells of type-H vessel to prevent alcoholic osteopenia.

Keywords: BMP2; Bmi-1; osteopenia; p16; proliferation; type-H vessel.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Aging
Animals
Bone Diseases, Metabolic* / metabolism
Cell Proliferation
Endothelial Cells*
Humans
Infant
Mice
Osteogenesis
X-Ray Microtomography